All products have special prices for bulk purchase, please contact for more details if required.

Explore Versatile Options with Our Glycerol-Free and Lyophilized Variants

Delight in the flexibility of our product offerings, now available in a glycerol-free version, meticulously crafted to seamlessly integrate with your freeze-drying systems. 

Additionally, we present a lyophilized version, a practical solution ensuring stable transportation at room temperature, thereby notably mitigating shipping costs, especially during long-distance transits.

Cat. No.: 5DAK-10 (for 10T)

Cat. No.: 5DAK-50 (for 50T)

Description

5' DNA Adenylation Kit is a convenient and efficient enzymatic method for adenylation modification of the 5' end of single-stranded DNA (ssDNA).

The ssDNA modified with a 5' adenylation generated using this kit is commonly used for the ligation of adapters to RNAs with a 3' hydroxyl end, such as miRNAs, or single-stranded DNAs with a 3' hydroxyl end in cloning, high-throughput sequencing library preparation, PCR detection, and other applications.

To perform the adenylation modification reaction using this kit, you will need Adenylase, ATP, and the 5'-phosphorylated single-stranded DNA to be modified with adenylation.

The adenylation efficiency of this kit is high, typically achieving over 95%. When performing adenylation modification on 5'-phosphorylated single-stranded DNA, regardless of whether the 3' end of the DNA is blocked or not, high yields of adenylation products can be obtained. Typically, this kit can convert over 95% of the 5'-phosphorylated DNA (pDNA) into adenylation DNA (AppDNA). Therefore, the adenylation products usually do not require gel purification and can be directly ethanol-precipitated for further concentration before use in subsequent ligation reactions.

This product utilizes 5'-phosphorylated single-stranded DNA as a substrate, and the adenylase enzyme hydrolyzes ATP into AMP and PPi. AMP is then transferred to the 5'-phosphate group of the single-stranded DNA, resulting in adenylation-modified single-stranded DNA and the preparation of adenylation adapters (linkers).

This product can also be used for adenylation modification of the 5' end of 5'-phosphorylated single-stranded RNA. Depending on the specific application, suitable experimental steps can be selected. Additionally, it is necessary to prepare RNase Inhibitor and DEPC water separately.

The Adenylase included in this product is derived from a thermophilic archaeon and expressed in Escherichia coli.

Advantages

The adenylation modification of 5'-phosphorylated single-stranded DNA or RNA can be completed in a single step, making it more convenient compared to traditional chemical methods. The conversion efficiency of the substrate is over 95%, and there is no need for gel purification; ethanol precipitation or similar methods are sufficient for subsequent reactions. The reaction can be performed at a higher temperature, such as 65°C, effectively avoiding interference from secondary structures on the adenylation modification of the 5' end. It is suitable for reaction systems with substrate quantities at the pmol level and can be easily scaled up to reaction systems with substrate quantities at the µmol level.

Precautions

• 5'-phosphorylation of the substrate single-stranded DNA or RNA is necessary, while the 3' end can be modified with amino groups or other blocking agents, or it can be left unmodified.
• The adenylation reaction system provided in this kit can be scaled up proportionally without significantly affecting the yield of adenylation products.
• The optimal reaction temperature for the Adenylase provided in this kit is 65°C. At 25°C, deadenylation may occur. Therefore, it is recommended to incubate at 85°C for 5 minutes to inactivate the Adenylase after the reaction is complete. If the Adenylase is not inactivated, subsequent exposure to room temperature conditions may result in a decrease in the adenylation rate.
• This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in ordinary residential areas.
• For your safety and health, please wear laboratory attire and disposable gloves when handling this product.

Storage

The minimum shelf life is 1 year at -20°C.